Using 454 technology for long-PCR based sequencing of the complete mitochondrial genome from single Haemonchus contortus (Nematoda).
Using 454 technology for long-PCR based sequencing of the complete mitochondrial genome from single Haemonchus contortus (Nematoda).
复制标题
使用454技术从单个Haemonchus contortus（Nematoda）的完整线粒体基因组进行长PCR测序。
DOI：
10.1186/1471-2164-9-11
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发表时间：
2008-01-11
期刊：
影响因子：
4.4
通讯作者：
Gasser, Robin B.
中科院分区：
文献类型：
--
作者：
Jex, Aaron R.;Hu, Min;Littlewood, D. Timothy J.;Waeschenbach, Andrea;Gasser, Robin B.
线粒体（mt）基因组是一个丰富的分子标记来源，在群体遗传学、系统学、流行病学和生态学等领域有着广泛的应用。在本研究中，我们使用454技术（或GS 20，大规模平行皮升反应器平台），以确定完整的mt基因组的螺旋血矛线虫（线虫纲：毛圆线虫科），寄生虫的重大农业，兽医和经济意义。我们验证这种方法通过与公开可用的表达序列标签（EST）和基因组调查序列（GSS）数据集的MT序列进行比较。采用454技术，利用来自单个成年雄性的基因组DNA（~20-40 ng），从长PCR扩增模板直接对捻转血矛线虫的完整mt基因组进行测序。组装单个重叠群，并与从公开可用的EST（NemBLAST）和GSS数据集挖掘的mt序列进行比较。比较结果表明，454技术平台对于线虫富含AT的mt基因组测序是可靠的。捻转血矛线虫mt基因组全长14，055 bp，AT含量高（78.1%）。与迄今为止研究的其他嗜色线虫一致，H. contortus包含36个基因（12个蛋白质编码，22个tRNA，rrnL和rrnS），并在结构，大小和基因排列的特征以前的圆线虫的成员相似。本研究证明了454技术用于从微量DNA中快速确定mt基因组序列的实用性，并揭示了当前可用于挖掘的数据库中的大量mt基因组数据。该方法为线虫和其他生物mt基因组的高通量测序提供了一个新的平台。
Mitochondrial (mt) genomes represent a rich source of molecular markers for a range of applications, including population genetics, systematics, epidemiology and ecology. In the present study, we used 454 technology (or the GS20, massively parallel picolitre reactor platform) to determine the complete mt genome of Haemonchus contortus (Nematoda: Trichostrongylidae), a parasite of substantial agricultural, veterinary and economic significance. We validate this approach by comparison with mt sequences from publicly available expressed sequence tag (EST) and genomic survey sequence (GSS) data sets. The complete mt genome of Haemonchus contortus was sequenced directly from long-PCR amplified template utilizing genomic DNA (~20–40 ng) from a single adult male using 454 technology. A single contig was assembled and compared against mt sequences mined from publicly available EST (NemBLAST) and GSS datasets. The comparison demonstrated that the 454 technology platform is reliable for the sequencing of AT-rich mt genomes from nematodes. The mt genome sequenced for Haemonchus contortus was 14,055 bp in length and was highly AT-rich (78.1%). In accordance with other chromadorean nematodes studied to date, the mt genome of H. contortus contained 36 genes (12 protein coding, 22 tRNAs, rrnL and rrnS) and was similar in structure, size and gene arrangement to those characterized previously for members of the Strongylida. The present study demonstrates the utility of 454 technology for the rapid determination of mt genome sequences from tiny amounts of DNA and reveals a wealth of mt genomic data in current databases available for mining. This approach provides a novel platform for high-throughput sequencing of mt genomes from nematodes and other organisms.
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影响因子：
4
作者：
Hu, M;Chilton, NB;Gasser, RB
通讯作者：
Gasser, RB
影响因子：
14.8
作者：
Burger, Gertraud;Lavrov, Dennis V.;Lang, B. Franz
通讯作者：
Lang, B. Franz
影响因子：
10.7
作者：
Blouin, MS;Yowell, CA;Dame, JB
通讯作者：
Dame, JB
影响因子：
2.1
作者：
Keddie, EM;Higazi, T;Unnasch, TR
通讯作者：
Unnasch, TR
影响因子：
64.8
作者：
Leininger, S.;Urich, T.;Schleper, C.
通讯作者：
Schleper, C.